The biological elimination of therapeutic 131I in patients with differentiated thyroid cancer (DTC), post total or near-total thyroidectomy, was compared after withholding levothyroxine suppression against administration of recombinant human thyrotropin without stopping levothyroxine. In 163 patients (group G1) levothyroxine was withheld before 131I therapy: in 138 patients the tumor was limited to the thyroid bed (group G1.1) and in 25 patients metastases were present (group G1.2). A second group of patients (G2; n = 28) received 131I therapy after administration of recombinant human thyrotropin without stopping levothyroxine. Mean retained 131I activity (as a percentage of the administered dose) was 5%-29% (group G1.1), 20%-43% (group G1.2) and 1%-17% (group G2). The effective half-life of 131I was 0.59-0.69 days (group G1.1), 0.87-1.22 days (group G1.2) and 0.38-0.44 days (group G2). In conclusion, the use of recombinant human thyrotropin to prepare patients with thyroid cancer for therapy with 131I shortens its effective half-life and reduces its retained activity compared to preparation with discontinuation of levothyroxine suppression.